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91.
Anna Fietta Guido Grandi Massimo Malcovati Gianna Valentini Vittorio Sgaramella Antonio G. Siccardi 《Plasmid》1981,6(1):78-85
Plasmids S-a and Rts1 suppress the galactose-sensitive phenotype of galE mutants of Escherichia coli K-12, giving rise to both galactose-fermenting and nonfermenting strains. Fermenting strains produce normal inducible UDP-galactose epimerase. Plasmids extracted from either a fermenting or a nonfermenting strain are indistinguishable when examined by either measurements of length of relaxed circular molecules by electron microscopy or electrophoretic pattern of restriction endonuclease digestion products. The phenomenon could be explained by reversible recombination between a plasmid-borne epimerase gene and homologous chromosomal sequences. 相似文献
92.
p-Hydroxyphenylglyoxal reacts with arginine residues in proteins to give a single product which can be quantitated spectrophotometrically. The reaction takes place under mild conditions, pH 7–9 and 25°C. Under these conditions up to complete modification of Nα-citraconyl-l-arginine was obtained within 60 min with less than 5% modification of other common amino acid side chains. The extent of modification in a protein can be determined at 340 nm using the molar absorption coefficient of 1.83 × 104m?1 cm?1 for the product at pH 9.0 and 25°C following removal of excess reagent by gel filtration. Several proteins, previously shown to have essential arginines, were modified by p-hydroxyphenylglyoxal and the losses in arginines were determined spectrophotometrically. These results were in close agreement with those of previous investigators. Rhea ovomucoid, a glycoprotein without arginines but containing an essential lysine, was relatively unaffected. 相似文献
93.
An informative family, in which parents shared HLA-Dw and Ia-like DRw (Ly-Li) antigens, was used to produce PLTs between members either phenoidentical for both Dw and DRw determinants or incompatible for Dw specificities only. These PLTs were restimulated by members of the family: two PLTs, although in DRw identity, reacted against members of the family bearing one maternal (c) and/or one paternal (a) haplotype. A third PLT also developed in DRw identity reacted with members bearing the other maternal (d) haplotype. Population studies with one of these PLTs did not show any correlation with Dw or DRw specificities. Family studies are in keeping, but do not demonstrate an HLA linkage. The data suggest that, along with the stimulating products (PLA) identical or closely related to the DRw determinants, other stimulating products (PLB), also probably HLA-linked, exist. Furthermore, one of the PLTs was produced without a primary MLR. 相似文献
94.
Antonio Baldini Dorothy A. Miller Viji Shridhar Mariano Rocchi Orlando J. Miller David C. Ward 《Chromosoma》1991,101(2):109-114
We have isolated an alpha satellite DNA clone, pG3.9, from gorilla DNA. Fluorescence in situ hybridization on banded chromosomes under high stringency conditions revealed that pG3.9 identifies homologous sequences at the centromeric region of ten gorilla chromosomes, and, with few exceptions, also recognizes the homologous chromosomes in human. A pG3.9-like alphoid DNA is present on a larger number of orangutan chromosomes, but, in contrast, is present on only tow chromosomes in the chimpanzee. These results show that the chromosomal subsets of related alpha satellite DNA sequences may undergo different patterns of evolution.by J.B. Rattner 相似文献
95.
Bonaventura Ruiz-Montasell F. Javier Casado Antonio Felipe Marçal Pastor-Anglada 《The Journal of membrane biology》1992,128(3):227-233
Summary The characteristics of uridine transport were studied in basolateral plasma membrane vesicles isolated from rat liver. Uridine was not metabolized under transport measurement conditions and was taken up into an osmotically active space with no significant binding of uridine to the membrane vesicles. Uridine uptake was sodium dependent, showing no significant stimulation by other monovalent cations. Kinetic analysis of the sodium-dependent component showed a single system with Michaelis-Menten kinetics. Parameter values were K
M 8.9
m and V
max 0.57 pmol/mg prot/sec. Uridine transport proved to be electrogenic, since, firstly, the Hill plot of the kinetic data suggested a 1 uridine: 1 Na+ stoichiometry, secondly, valinomycin enhanced basal uridine uptake rates and, thirdly, the permeant nature of the Na+ counterions determined uridine transport rates (SCN– > NO
3
–
> Cl– > SO
4
2–
). Other purines and pyrimidines cis-inhibited and trans-stimulated uridine uptake.This work has been partially supported by grant PM90-0162 from D.G.I.C.Y.T. (Ministerio de Educación y Ciencia, Spain). B.R.-M. is a research fellow supported by the Nestlé Nutrition Research Grant Programme. 相似文献
96.
Raúl Alva J. Antonio Lugo-R Eduardo Arzt Jorge Cerbón Blanca E. Rivera Mireya Toro Sergio Estrada-O 《Journal of bioenergetics and biomembranes》1992,24(1):125-129
Nigericin is a monocarboxylic polyether molecule described as a mobile K+ ionophore unable to transport Li+ and Cs+ across natural or artificial membranes. This paper shows that the ion carrier molecule forms complexes of equivalent energy demands with Li+, Cs+, Na+, Rb+, and K+. This is in accordance with the similar values of the complex stability constants obtained from nigericin with the five alkali metal cations assayed. On the other hand, nigericinalkali metal cation binding isotherms show faster rates for Li+ and Cs+ than for Na+, K+, and Rb+, in conditions where the carboxylic proton does not dissociate. Furthermore, proton NMR spectra of nigericin-Li+ and nigericin-Cs+ complexes show wide broadenings, suggesting strong cation interaction with the ionophore; in contrast, the complexes with Na+, K+, and Rb+ show only clear-cut chemical shifts. These latter results support the view that nigericin forms highly stable complexes with Li+ and Cs+ and contribute to the explanation for the inability of this ionophore to transport the former cations in conditions where it catalyzes a fast transport of K+>Rb+>Na+.Part of the results of this paper were presented at the 14th International Congress of Biochemistry in Prague, Czechoslovakia. 相似文献
97.
Mario Díaz Antonio Lorenzo 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1992,162(2):189-196
Summary Bioelectrical parameters and unidirectional sodium and chloride fluxes were measured under voltageclamp conditions in groups of lizards submitted to single or chronic aldosterone treatment. Both acute (AT) and chronic (CT) treatment induced significant increases in the short-circuit current (I
sc), as well as in the mucosa-to-serosa (J
m-s
Na
) and net sodium flux (J
net
Na
). In AT tissues, aldosterone did not change net chloride flux (J
net
Cl
) but did so in CT tissues. Amiloride reduced the aldosterone-increased I
sc in AT and CT tissues, inhibited J
net
Na
in AT tissues and abolished it in CT colons. J
net
Cl
was also reduced by the diuretic in the group of AT colons, whereas no changes were observed in the CT tissues. Addition of luminal DIDS reduced Na+ absorption and totally inhibited Cl- absorption in the AT tissues, but did not change I
sc. However, in CT tissues neither Na+ nor Cl- transport were affected by DIDS. A good relationship between I
sc and J
m-s
Na
was apparent after DIDS treatment in AT tissues. In this group, simultaneous addition of DIDS and amiloride totally abolished J
net
Na
and reduced I
sc to untreated control values. Addition of serosal ouabain abolished I
sc and Na+ absorption in AT and CT colons, but Cl- absorption was only altered in AT tissues. These results support the hypothesis that aldosterone induces an electrogenic, amiloride-sensitive sodium absorption, and in a dose-dependent fashion suppresses electroneutral NaCl absorption in the lizard colon.Abbreviations AT
acutely treated
- CT
chronically treated animals
- DIDS
4-4-diisothiocyanatostibene-2-2-disulfonic acid
- DMSO
dimethylsulphoxide
-
G
t
tissue conductance
-
I
sc
short circuit current
- PD
transepithelial potential difference
- SITS
4-acetamido-4-isothiocyanatostilbene-2-2-disulfonic acid
- UC
untreated controls
Preliminary results of this paper were presented at the X
th meeting of the European Intestinal Transport Group (EITG), Askov Hojskole, Denmark, 16–19 September 1990 相似文献
98.
99.
Following i.c.v. (intracerebral ventricular) injections ofd,l-[3H]pipecolic acid (PA), it is reabsorbed from the ventricles and redistributed to various brain regions. The highest accumulation is found in three brain regions ipsilateral to the injection site, hippocampus, neocortex, striatum, and in the diencephalon. Following preloading in vivo, the radioactivity is released from hippocampus slices in the perfusion medium after depolarization induced by high K+. During perfusion with a Ca++ free medium containing EGTA, a significant reduction of release is observed.The radioactivity ofd,l-[3H]PA in the brain shows a more rapid phase of decrease from 0 to 2 hours and a slower phase from 2 to 5 hours. At 5 hours, only 28% radioactivity, represented mainly by PA, is left in the brain. Kidney secretion represents the major route of elimination of the injected PA. The presence of -aminoadipic acid both in brain and urine was observed. Probenecid (200 mg/kg) significantly increases the accumulation of i.c.v. injectedd,l-[3H]PA in brain and kidney. The presence of a regional accumulation of PA in certain brain regions, its metabolism in brain, its enhanced retention following probenecid administration and its Ca++ dependent release following high K+ stimulation, all constitute indirect evidence for a neuronal localization of this brain endogenous iminoacid. 相似文献
100.
The catalytic mechanism of cytochrome P-450. Spin-trapping evidence for one-electron substrate oxidation 总被引:3,自引:0,他引:3
O Augusto H S Beilan P R Ortiz de Montellano 《The Journal of biological chemistry》1982,257(19):11288-11295
Cytochrome P-450 is destroyed during catalytic oxidation of several 4-substituted 3,5-bis(ethoxycarbonyl)-2,6-dimethyl-1,4-dihydropyridine substrates. A qualitative correlation has been found between the ability to destroy cytochrome P-450 and the stability of the 4-substituent as a radical. Destruction of the enzyme by the 4-ethyl (DDEP), 4-propyl, and 4-isobutyl analogues is due to transfer of the 4-alkyl group from the substrate to a nitrogen of the prosthetic heme, a process which gives rise to isolable N-alkylprotoporphyrin IX derivatives. Little enzyme destruction is observed when the 4-alkyl group is of low radical stability (methyl, phenyl) and good destruction, but no isolable heme adducts when the 4-substituent is of very high radical stability (isopropyl, benzyl). Spin-trapping studies have established that the 4-ethyl group in DDEP is lost as a radical as a result of oxidation by cytochrome P-450. Of three commonly used spin traps, only alpha-(4-pyridyl-1-oxide) N-tert-butylnitrone was found suitable for such studies. The other spin traps, 5,5-dimethyl-1-pyrroline-N-oxide and alpha-phenyl N-tert-butylnitrone, were found to be ineffective, the latter because it strongly inhibits cytochrome P-450. Hydrogen peroxide formed in situ can support a part of the cytochrome P-450-catalyzed ethyl radical formation and DDEP-dependent self-inactivation. The results provide persuasive evidence that oxidation of the nitrogen in DDEP by cytochrome P-450 proceeds in one-electron steps. Cytochrome P-450 may thus function, at least with certain substrates, as a one-electron oxidant. 相似文献